Method for bioremediating undetonated explosive device

ABSTRACT

Technology for in situ remediation of undetonated explosive device. An explosive device contains an explosive material in close proximity with microorganisms capable of metabolizing the explosive material that are either mobile or temporarily deactivated by freeze drying. Examples include  Pseudomonas  spp.,  Escherichia  spp.,  Morganella  spp.,  Rhodococcus  spp.,  Comamonas  spp., and denitrifying microorganisms. A self-remediating explosive mixture includes an explosive material intermixed with microorganisms. Joined with an explosive device is a bioremediation apparatus that contains microorganisms and prevents contact between microorganisms and explosive material in the explosive device using a barrier that is actuated to release the microorganisms by mechanical, electrical, or chemical mechanisms. If the explosive device fails to detonate, remediation by microorganisms includes both disabling of the explosive material and detoxification of resulting chemical compositions.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional application of U.S. patent applicationSer. No. 09/666,073 that was filed on Sep. 19, 2000 (hereinafter “theParent Application”), and that issued as U.S. Pat. No. 6,644,200 on Nov.11, 2003. The Parent Application is a divisional application of U.S.patent application Ser. No. 08/743,460 that was filed on Oct. 18, 1996(hereinafter “the Grandparent Application”), and that issued as U.S.Pat. No. 6,120,627 on Sep. 19, 2000. The Grandparent Application is acontinuation-in-part application of both U.S. patent application Ser.No. 08/658,104 that was filed on Jun. 4, 1996, now abandoned which is acontinuation-in-part of U.S. patent application Ser. No. 08/560,074 thatwas filed on Nov. 17, 1995, now abandoned and U.S. patent applicationSer. No. 08/687,092 that was filed Jun. 4, 1996, now abandoned which isa continuation-in-part application of U.S. patent application Ser. No.08/560,102 that was filed on Nov. 17, 1995 now abandoned.

This application discloses subject matter related to that disclosed inU.S. Pat. No. 5,763,815 that issued on Jun. 9, 1998, from U.S. patentapplication Ser. No. 658,995 filed on Jun. 4, 1996, and which is acontinuation-in-part of U.S. patent application Ser. No. 560,527 thatwas filed on Nov. 17, 1995, and subject matter related to that disclosedin U.S. Pat. No. 5,736,669 that issued on Apr. 7, 1998, from U.S. patentapplication Ser. No. 658,142 that was filed Jun. 4, 1996, and which is acontinuation-in-part of U.S. patent application Ser. No. 560,074, filedon Nov. 17, 1995.

This application also discloses subject matter related to that disclosedin U.S. Pat. No. 6,660,112 that issued on Dec. 9, 2003, from U.S. Pat.application Ser. No. 666,020 that was filed on Sep. 19, 2000, and thatis related to the same applications and in the same manner as those setforth above for the Parent Application.

BACKGROUND

The present invention is directed to systems, apparatus, and methods forremediating explosives. More particularly, the present invention isdirected to the remediation of explosives which have not detonated.

Explosive charges are inherently dangerous in a number of respects.

Inadvertent detonation poses risks of severe personal injury or death,as well as of substantial property destruction and consequential losses.Explosive charges are, in addition, comprised of material substances,which even when not consolidated in a shape capable of performing as adetonatable explosive charge, may be toxic and thus potentiallyinjurious to human health and to complex as well as simple plant andanimal life.

Explosive charges that are not securely stored in a supervised manner,or isolated from the environment and from indiscriminate access by humanand animal life forms, thus present both safety and environmentalhazards.

Such hazards are pointedly apparent where an explosive charge fails todetonate after the explosive charge has been installed for that purposeduring activities pertaining to mining, construction, or to seismicsurveying. Fortunately, installed explosive charges that do not detonateas planned are usually locatable and often recoverable through theexpenditure of reasonable efforts and without safety risks to personnel.On the other hand, there do routinely arise circumstances in whichundetonated explosive charges of this type are not recovered or simplycannot be recovered. Then, the risks are present that the undetonatedexplosive charge could at some subsequent time be detonatedinadvertently or become a source of potentially harmful contaminants.

As an example, seismic survey data used to ascertain the nature ofsubsurface ground structures is routinely obtained by recording andanalyzing shock waves that are propagated into the ground and producedby detonating explosive charges. The shock waves are then monitoredduring transmission through the ground. In this role, such seismiccharges are usually utilized in large sets, installed as an array ofindividual seismic charges at widely disbursed locations. The seismiccharges are interconnected with detonation equipment for remotedetonation, either simultaneously or in sequence.

Seismic charges for such surveys can be detonated either above or belowthe surface of the ground. In either case, it is not uncommon that atleast one of any set of such seismic charges does not detonate asintended. Such failures may be caused by defects in the explosive chargeitself, by damage caused during installation, by faulty detonationequipment, or by the failure of personnel in the field to make effectiveinterconnections between detonation equipment and each seismic charge inthe installed set.

When a seismic charge installed above the ground fails to detonate asintended, it is usually possible to locate and safely recover theundetonated seismic charge. Nonetheless, circumstances do exist wherethe detonation of a set of seismic charges installed above the grounddislocates one of the undetonated seismic charges in the set, directingthat undetonated seismic charge into a terrain in which the chargecannot be located or cannot be recovered easily. Responsible seismiccrews naturally are trained to exercise all reasonable efforts torecover undetonated seismic charges that are located on the surface ofthe ground, but even the most rigorously indoctrinated and enthusiasticseismic personnel cannot guarantee that all undetonated seismic chargesinstalled above the ground are ultimately recovered.

Aside from the human factor involved, the intervention of severe weatherconditions, such as sandstorms, blizzards, tornadoes, or hurricanes, canimpede efforts to recover undetonated seismic explosives. Some suchweather conditions offer the prospect of even altering the terrain,thereby burying the undetonated seismic charge temporarily or for asubstantial duration. Floods can cover the seismic survey site, removingor obscuring undetonated seismic charges. In the extreme, geologicalsurface changes, such as mudslides, rockfalls, and fissures caused byearthquakes, by heavy weather, or even by seismic survey activityitself, can preclude the recovery of undetonated seismic charges, andeven obscure the understanding that any seismic charge has failed todetonate.

The safety risks and environmental hazards posed by loose, undetonatedexplosive charges will be present where any undetonated seismic chargeremains unrecovered after the detonation of the set of seismic chargesof which it was a part.

The likelihood that an undetonated seismic charge will be abandoned isgreatest, however, relative to the conduct of seismic survey activitybased on the detonation of seismic charges installed below the surfaceof the ground. In such subsurface seismic detonation activity, a seriesof deep boreholes are drilled into the earth or rock at predeterminedlocations that are intended to maximize the data to be derived from theshock waves promulgated from the detonation of the seismic charges. Aseismic charge is placed at the bottom of each borehole and then shut inthe borehole in a relatively permanent manner using a concrete or asealing compound, such as bentonite. The balance of the borehole is thenbackfilled with loose soil and rock, a process which alone accounts forthe majority of failed seismic detonations. Backfill materials have anunderstandable tendency to break the detonating cord leg wires ornon-electric transmission line that interconnects the installed seismiccharge at the bottom of the borehole with detonating equipment locatedabove the ground. If a seismic charge installed below the ground failsto detonate, the easy removal of the undetonated seismic charge isseriously impeded by yards of backfill and the cured concrete or sealingcompound in which the seismic charge was embedded at the bottom of theoriginal borehole. Removing such an installed seismic charge byreexcavating the original borehole or by digging around the originalborehole to avoid the sealing compound is extremely laborious and timeconsuming, potentially unsafe, and in many circumstances virtuallyimpossible.

Thus, in conducting seismic survey activities, particularly seismicsurvey activities involving the detonation of seismic charges below thesurface of the ground, undetonated seismic charges are regularlyabandoned in the field. Frequently, even the precise location ofundetonated seismic charges cannot be pinpointed. The risks fromundetonated explosive charges installed in the ground endure for asubstantial time, usually exceeding the durability of ground surfacewarning signs, fencing, or the continued possession and control ofaccess to the site by an original owner. Eventually, the pressure ofhuman population growth may render the site attractive for civil orindustrial activities that would not be consistent with buriedundetonated explosive charges.

The associated dangers include first that of an accidental detonation atsome future time. Less dramatic, but certainly of longer duration, arerisks presented by the material substance of those undetonated charges.Once released from the confines of the casing of an explosive assembly,the explosive material therein may cease to present any risk ofexplosion. This type of release of explosive materials can occur throughcorrosion of the casing through the action of ground water, the fractureof the casing during careless installation, or the shifting of theground structure at the location at which the undetonated seismic chargewas abandoned. In due course, the prolonged effect of these forces incombination with surface erosion or subsurface fluid migration candisburse over a large area the material of a fractured explosive charge.That material may constitute a potentially problematic contaminant. Evenif detected, remedial activities may be required to contain and possiblyeliminate the contaminant.

Nonetheless, no practical methods exist for reliably remediating therisks posed by undetonated explosive charges, particularly where thoseundetonated explosive charges are originally installed below the surfaceof the ground.

SUMMARY OF THE INVENTION

It is thus the broad object of the present invention to protect publichealth and safety from risks arising from incidents of abandonedundetonated explosive charges.

Accordingly, it is a related object of the present invention toeliminate the possibility of detonation of abandoned explosive charges.

It is a complementary object of the present invention to reduce thelikelihood that abandoned undetonated explosive charges will contributeto environmental pollution.

Thus, it is a specific object of the present invention to provideapparatus, systems, and methods for remediating in situ any installedexplosive charge that fails to detonate as intended.

It is a particular object of the present invention to provide suchapparatus, systems, and methods as are capable of reliably and safelyremediating an undetonated explosive charge abandoned in the ground.

Yet a further object of the present invention is to provide suchapparatus, systems, and methods as are capable of remediating anundetonated explosive charge, even if the location of the explosivecharge cannot be ascertained with any degree of certainty.

These and other objects and features of the present invention willbecome more fully apparent from the following description and appendedclaims, or will be appreciated by the practice of the invention.

To achieve the foregoing objects, and in accordance with the inventionas embodied and broadly described herein, apparatus, systems, mixturesand methods are provided that remediate in situ an undetonated explosiveutilizing the biological activity of microorganisms.

In one form, an apparatus incorporating teachings of the presentinvention includes a quantity of explosive material and microorganismsthat are disposed in sufficient proximity to the quantity of theexplosive material that the microorganisms can initiate bioremediationof the explosive material when the microorganisms are mobile. Similarly,an explosive mixture is formed by intermixing the microorganisms and theexplosive material. The explosive apparatus preferably has a shell thatenables water to flow through the shell to contact the explosivematerial. The shell may for example have an open end, have holes or bewater permeable.

The apparatus or mixture may also further comprise a mobilization meansfor mobilizing the microorganism to contact the explosive material. Themobilization means enables the microorganisms to initiate bioremediationof the explosive material or to continue bioremediating the explosivematerial. The terms “mobile” and “mobility” refer to the ability of themicroorganisms to move, to be carried by the movement of a liquid, to bedistributed to the explosive material or to be unrestricted in movementby a barrier that previously confined the microorganisms such that afterthe barrier is removed the microorganisms can contact the explosivematerial. The term “active” refers to the state of the microorganismswherein the microorganisms can bioremediate explosives.

An example of a mobilization means that is useful with an explosiveapparatus includes a rigid mechanical structure having a barrier toprevent contact of the microorganisms with the explosive material untilthe barrier is removed and the microorganisms are mobilized to contactthe explosive material. The barrier can be removed by a mechanism thatis mechanical, electrical and/or chemical. Other examples ofmobilization means which can be utilized with an explosive apparatus oran explosive mixture include a mobilizing liquid such as water or aliquid with nutrients, a sufficient degree of porosity in the explosivematerial or the explosive mixture, and surfactants in the explosivematerial or mixture.

The microorganisms can be mobile or deactivated. Examples of deactivatedmicroorganisms include microorganisms that have been dehydrated by airdrying or by being lyophilized. The microorganisms are preferably freezedried to increase the survivability of the microorganisms during theforming process wherein the explosive material and microorganisms arecombined. More specifically, it is desirable to heat the explosivematerial to increase the moldability of the explosive material and toenable the microorganisms and explosive material to be easilyintermixed; however, the heat can be lethal to the microorganisms as themicroorganisms are placed or mixed in the explosive material.Accordingly, the microorganisms have preferably been prepared such thatthe microorganisms can be characterized in that the microorganisms aresufficiently resistant to heat that a significant portion of themicroorganisms survive the intermixing or placement process even whenthe process occurs at a temperature of about 100° C.

The microorganisms can be disposed in close proximity to the explosivematerial or dispersed within the explosive material in many differentforms. The microorganisms can be in various aggregations such as inpellets or in capsules. The aggregations can also be added without anyprocessing of the microorganisms to form the microorganisms into aparticular distinct form. Accordingly, the microorganisms can be presentas a flake, granule, clump, powder or shard of a nutrient mediumcontaining microorganisms. Nutrients, in addition to the explosivematerial, are generally necessary for the microorganisms to survive andgrow. Binders are also often necessary and organic binders arepreferred. Depending on the binder or nutrient utilized, one chemicalcan perform the function of both binder and nutrient. The thermalresistance of the microorganisms can also be increased by utilizingvarious thermal protection additives.

An example of a structure having a barrier to prevent contact of themicroorganisms with the explosive material until the barrier is removedis provided by a bioremediation apparatus in combination with anexplosive material. The bioremediation apparatus includes a storagemeans for releasably containing at least one type of microorganismcapable of degrading explosive materials. Stored distinctly therefrom inthe bioremediation apparatus is a reservoir means for releasablycontaining a liquid intended to be mixed with the microorganisms. Thestorage means is positioned proximate the reservoir means, usually in arelationship that is below the reservoir means in the anticipatedinstalled orientation of the inventive apparatus. The bioremediationapparatus further includes a first valve means for delivering the liquidfrom the reservoir means to the microorganisms in a storage means. Doingso causes mobilization of the microorganisms. This occurs when a firstvalve means is opened. The first valve means is at least partiallydisposed within the reservoir means.

Additionally, the bioremediation apparatus of the present inventioncomprises a second valve means for delivering hydrated microorganisms toan associated, undetonated explosive material. The second valve means isoperably linked to the first valve means and is at least partiallydisposed within the storage means.

The bioremediation apparatus is coupled in one embodiment of the presentinvention with an explosive apparatus that has an actuation means foropening the first valve means and the second valve means upon beingcoupled thereto. The actuation means for opening the valves can beachieved by either a mechanical or electrical mechanism. If theexplosive material in the explosive apparatus fails to detonate, theexplosive material will eventually be remediated by the action of themicroorganisms released from the associated storage means.

Ideally, the remediation occurs in two respects. The explosive isdisabled from inadvertent detonation. Subsequently, the materialcomposition of the explosive material is rendered relatively nonharmful.

In another embodiment of the invention, microorganisms are releasablycontained be gelatin, a substance that is self-effacing when contactedby microorganisms under favorable condition. For example, gelatin may beused to fabricate the first valve means that retains liquid in thereservoir mans of the bioremediation apparatus or the second valve meansthat retains the microorganism in the storage means of thebioremediation apparatus.

In yet another embodiment, microorganisms are applied directly to theexterior of the explosive material or to the shell of an explosiveapparatus.

BRIEF DESCRIPTION OF THE DRAWINGS

In order that the manner in which the above-recited and other advantagesand objects of the invention are obtained, a more particular descriptionof the invention briefly described above will be rendered by referenceto specific embodiments thereof, which are illustrated in the appendeddrawings. Understanding that these drawings depict only typicalembodiments of the invention and are not, therefore, to be considered tobe limiting of the scope of the invention, the present invention will bedescribed and explained with additional specificity and detail throughthe use of the accompanying drawings in which:

FIG. 1 is a perspective view of a first embodiment of a bioremediationapparatus incorporating teachings of the present invention.

FIG. 2 is a perspective view in partial break-away of the bioremediationapparatus of FIG. 1 in the process of being coupled with an explosiveapparatus in accordance with teachings of the present invention.

FIG. 3 is a cross-sectional elevation view of the bioremediationapparatus and the explosive apparatus illustrated in FIG. 2 taken alongsection line 3—3 shown therein.

FIG. 4 is a perspective view in partial break-away of the bioremediationapparatus and the explosive apparatus of FIG. 2 immediately uponbecoming fully coupled.

FIG. 5 is a cross-sectional elevation view of the bioremediationapparatus and the explosive apparatus illustrated in FIG. 4 taken alongsection line 5—5 shown therein.

FIG. 6 is a cross-sectional elevation view like that of FIG. 5illustrating the bioremediation apparatus and explosive apparatus inFIG. 4 at a time subsequent to that illustrated in FIG. 5 at which theexplosive material in the explosive apparatus illustrated is contactedby hydrated microorganisms.

FIG. 7 is a partial cross-sectional elevation view like that of FIG. 4,but of a second embodiment of a bioremediation apparatus immediatelyincorporating teachings of the present invention immediately uponbecoming fully coupled with an explosive apparatus.

FIG. 8 is a partial cross-sectional elevation view like that of FIG. 6,but of the second embodiment of the bioremediation apparatus and theexplosive apparatus illustrated in FIG. 7 at a time subsequent to thatillustrated in FIG. 7 at which the explosive material in the explosiveapparatus illustrated is contacted by hydrated microorganisms.

FIG. 9 is a cross-sectional elevation view of a third embodiment of anexplosive apparatus that utilizes an electrical mechanism to control themobilization of the microorganisms.

FIG. 10 is a cross-sectional elevation view of a fourth embodiment of anexplosive apparatus comprising a pellet of microorganisms intermixed inthe explosive material.

FIG. 11 is a partial cross-sectional elevation view of a fifthembodiment of an explosive apparatus which comprises an encapsulatedpellet.

FIG. 12 is a partial cross-sectional elevation view of a sixthembodiment of an explosive apparatus which comprises an encapsulatedsuspension of microorganisms.

FIG. 13 is a partial cross-sectional elevation view of a seventhembodiment of an explosive apparatus which comprises shards of moistnutrient wafers containing microorganisms.

FIG. 14 is a partial cross-sectional elevation view of an eighthembodiment of an explosive apparatus which comprises a powder ofmicroorganisms dispersed on top of the explosive material.

FIG. 15 is a partial cross-sectional elevation view of a ninthembodiment of an explosive apparatus which depicts a chamber in theexplosive material containing a suspension of microorganisms.

FIG. 16 is a partial cross-sectional elevation view of a tenthembodiment of an explosive apparatus which comprises clumps ofmicroorganisms within the shell of the apparatus.

DETAILED DESCRIPTION OF THE INVENTION

The present invention pertains to systems, apparatus, and methods forthe in situ remediating of undetonated explosive charges. Themethodology employs at least one type of microorganism that is capableof digesting an explosive material.

According to the teachings of the present invention, an explosive chargeto be installed, for example by being buried in the ground, is so housedin a casing with the microorganisms. If the explosive charge fails todetonate, the explosive charge can then reliably be left undisturbed,and the microorganisms will digest or degrade the explosive materialinvolved. Preferably, the explosive will be thereby both disabled fromdetonation and detoxified.

The terms “remediate” and “remediation” are used in the specificationand the appended claims to refer generally to the conversion ortransformation of an explosive material which is detonatable by shock orheat into a different chemical material which is less explosive ornonexplosive. The terms “bioremediate” and “bioremediation” are used torefer to remediation effected by the action of microorganisms. Thepresent invention is thus one intended to bioremediate explosivematerials.

The present invention has demonstrated an immediate utility relative tohighly explosive materials, such as trinitrotoluene (TNT),pentaerythritol tetranitrate (PETN), cyclotrimethylene trinitramine(RDX), and cyclotetramethylene tetranitramine (HMX). These are typicallyutilized in seismic charges.

The term “bioremediable explosive” is used in the specification and theappended claims to refer to any explosive material which can beconverted into a less explosive or nonexplosive material by the actionof microorganisms, whether or not such microorganisms are explicitlydisclosed herein. The highly explosive materials listed above are thusbioremediable explosives, since it has been demonstrated that at leastthe examples of microorganisms disclosed herein are capable ofconverting those high energy explosive materials into less explosive ornonexplosive materials.

Currently, on the basis exclusively of the examples of microorganismsdisclosed herein, known bioremediative explosives include at leastexplosives which are classified as organic nitroaromatics, organicnitramines, or organic nitric esters. Examples of organic nitroaromaticsinclude TNT, hexanitrostilbene (HNS), hexanitroazobenzene (NAB),diaminotrinitrobenzene (DATB), and triaminotrinitrobenzene (TATB).Examples of organic nitramines include RDX, HMX, nitroquanidine (NQ),and 2,4,6-trinitrophenylmethylnitramine (tetryl). Examples of organicnitric esters include PETN, nitroglycerine, and ethylene glycoldinitrate.

In one embodiment of the present invention, highly explosive materials,such as TNT and PETN, are converted through the action of microorganismsinto less explosive materials. These intermediate chemicals can then befully transformed into materials such as biomass and chemicals such asCO₂ and N₂. Optimally, the highly explosive materials are reducedaccording to the teachings of the present invention, first into lessexplosive intermediate chemicals or nonexplosive products. Theseintermediate chemicals can then be further transformed as needed intoconstituents which are either less explosive or less harmful ascontaminants in the environment to the health of humans, animals orplants than the intermediate chemicals may be. The final productresulting from the metabolizing action of the microorganisms will thusinclude any number of combinations of elements that originated in theexplosive material as constituted before the initiation of thebioremediation process.

The microorganisms comprise at least a first type of microorganism thatdisables or deactivates the explosive material by degrading theexplosive material into less explosive materials or nonexplosivematerials. The microorganisms may also further comprise a second type ofmicroorganism that further bioremediates any intermediate chemicalsresulting from the bioremediation action of the first type ofmicroorganism to fully bioremediate the explosive material intononexplosive materials.

Although any type of microorganism capable of converting explosivematerial into less harmful chemicals is considered to be within thescope of the present invention, examples of microorganisms that havebeen demonstrated to exhibit that capacity include the group consistingof Pseudomonas spp., Escherichia spp., Morganella spp., Rhodococcusspp., Comamonas spp., and dentrifying microorganisms. It is within thescope of the present invention to use any combination of theseparticular microorganisms, or of any other microorganisms that aredetermined to be capable of bioremediating explosive materials. SuitablePseudomonas spp. microorganisms include microorganisms in the groupconsisting of aeruginosa, fluorescens, acidovorans, mendocina, cepacia,and an unidentified type.

The present invention thus utilizes any of numerous different selectionsof microorganisms capable of degrading explosive materials in any ofvarious relative quantities. Each of these various selections ofmicroorganisms will for convenience hereinafter and in the appendedclaims be referred to as a “microorganism consortium.” In such amicroorganism consortium, one type of microorganism can advantageouslyreduce the explosive material to a particular intermediate chemical,such as azoaromatics, while that type or another type of microorganismmay then further reduce the azoaromatics or other intermediate chemicalsto carbon chains, CH₄, NH₃, and N₂. In one presently preferredembodiment, such a microorganism consortium utilizes all or some ofvarious of the microorganisms belonging to Pseudomonas spp., Escherichiaspp., Morganella spp., Rhodococcus spp., Comamonas spp., anddenitrifying microorganisms.

The bioremediation rate is an important variable in designing a systemthat is impacted by many factors. One factor that is closely related tothe bioremediation rate of explosive materials by the microorganisms isthe growth rate of the microorganisms. The growth rate of some speciesof microorganisms disclosed herein are logarithmic while others are onlylinear. Accordingly, the growth rate of the consortium depends on thetype of microorganisms utilized. Additionally, the growth rate of theconsortium of microorganisms depends on other factors, such as theavailability of nutrients. The growth rate of the consortium ofmicroorganisms can, however, be generally characterized as logarithmic.

A consortium of microorganisms within the scope of the present inventionwas deposited on May 23, 1996, with the American Type Culture Collection(hereinafter “ATCC”) in accordance with the provisions of the BudapestTreaty on the International Recognition of the Deposit Microorganismsfor the Purpose of Patent Procedure. The ATCC is located at 10801University Boulevard, Manassas, Va. 20110-2209 U.S.A. The depositedconsortium of microorganisms was assigned ATCC Designation No. 55784.For purposes of this disclosure, the microorganism consortium depositedwith the ATCC and designated ATCC Designation No. 55784 is herebyincorporated by reference.

The microorganism consortium deposited with the ATCC was obtained fromRichards Industrial Microbiology Laboratories, Inc. (hereinafter “RIML”)located at 55 East Center, Pleasant Grove, Utah 84062 U.S.A. Themicroorganism consortium is identified at RIML by Product No. RL-247.Accordingly, microorganisms sold as RL-247 by RIML under the tradenameRL-247 and assigned ATCC Designation No. 55784 are considered to bewithin the scope of the invention disclosed herein, whether or notconstituent microorganisms therein are explicitly identified to anydegree herein.

The microorganisms of the microorganism consortium are chosen for havinga demonstrated ability to metabolize and degrade explosive materials inany way that contributes to the disabling of the explosive material orto the detoxification of the chemical components thereof. Ifmicroorganisms are selected that are both aerobic and anaerobic,bioremediation will occur in shallow and exposed surface locations, aswell as in deep explosive boreholes. Ideally, the microorganismsselected for the microorganism consortium should be nonpathogenic andsurfactant-producing, as this enhances the digestive action of themicroorganism colony.

In one embodiment of a microorganism consortium chosen according to theteachings of the present invention, the Pseudomonas spp. are selectedfrom the group consisting of aeruginosa, fluorescens, acidovorans,mendocina, and cepacia. Any microorganisms of Pseudomonas spp. otherthan the microorganisms identified above are considered to be within thescope of the invention disclosed herein, provided that suchmicroorganisms perform any of the functions described above havingutility in the remediating of an explosive charge. Correspondingly, anymicroorganism is considered to be within the scope of the inventiondisclosed herein, provided the microorganism exhibits any utilityrelative to the bioremediating of explosive materials.

Thus, the disclosure and incorporation herein of the microorganismconsortium assigned ATCC Designation No. 55784 or the disclosure of themicroorganism consortium available from RIML under the tradename RL-247,are but examples of microorganism consortiums within the teachings ofthe present invention and are not limiting of the microorganisms thatmay be selected for inclusion in a microorganism consortium according tothe teachings of the present invention.

Various embodiments of explosives are set forth hereinbelow which areconfigured to enable microorganisms to bioremediate a quantity ofexplosive material. The microorganisms are disposed in sufficientproximity to the explosive material that the microorganisms initiatebioremediation of the explosive material when the microorganisms aremobilized.

The shelf lives of the explosive material and the microorganisms areincreased by delaying the bioremediation activity of the microorganismsat least until the explosive is ready to be utilized. Accordingly, thepreferred embodiments involve the use of microorganisms that aretemporarily immobilized or have been blocked from contact with theexplosive material until the explosive is to be positioned in the groundor after the explosive is in the ground. Configurations can also beutilized wherein the microorganisms are initially mobile when positionedrelative to the explosive material, thereby enabling the microorganismsto immediately initiate bioremediation.

The embodiments of the invention designed to delay the bioremediationactivity of the microorganisms until a set time utilize a mobilizationmeans for mobilizing the microorganisms to contact the explosivematerial. The mobilization means enables the microorganisms to initiatebioremediation or continue bioremediation of the explosive material. Anymobilizing means can be utilized including mechanisms which areprimarily mechanical, electrical, chemical or combinations thereof.

Examples of combinations of mechanical and chemical mechanisms utilizedto mobilize the microorganisms are provided by the embodiment in FIGS.1–6 and the embodiment in FIGS. 7–8. An embodiment is depicted in FIG. 9that utilizes electrical, mechanical and chemical mechanisms to mobilizethe microorganisms. In these embodiments, a rigid mechanical structurecontains the microorganisms in a relatively immobilized condition or atleast separate from the explosive material. Bioremediation of theexplosive material is initiated when a barrier between themicroorganisms and the explosive material is removed and themicroorganisms are in an adequate quantity of a liquid to enable themicroorganisms to be sufficiently mobile to flow into contact with theexplosive materials.

A first embodiment of an apparatus employing principles of the presentinvention is illustrated in FIG. 1 as an explosive bioremediationapparatus 10. Bioremediation apparatus 10 includes a casing 12 having atop end 14 and a bottom end 16. Casing 12 is preferably formed from amaterial which is water resistant and is capable of withstandingextremes of temperature.

A cap 18 is inserted into top end 14 of casing 12. Cap 18 is preferablyformed from a durable material that will withstand being driven down aborehole with a tamping pole. Cap 18 includes a cap top 20 and anexternal cap member 22 integrally extending from cap top 20 and havingcap threads 24. Cap 18 is secured about top end 14 of casing 12 byengaging cap threads 24 with end threads 26 that are formed on theexterior of top end 14. Cap 18 may include an internal cap member withan O-ring or a foam seal so configured and positioned as to engage topend 14 of casing 12. This increases the security of the seal produced.

Cap 18 is but one example of a structure capable of functioning as a capmeans for sealing the top end of a casing, such as casing 12. Anotherexample of a structure capable of performing the function of a cap meansaccording to the teachings of the present invention would be a casingwithout any external cap member, but rather having an internal capmember that is inserted into top end 14. Alternatively, bioremediationapparatus 10 could be provided with a structure that performs thefoundation of such a cap means but is integrally formed with casing 12.Any such cap structure that is integrally formed with casing 12 from aplastic material should be constructed to withstand the impacts andpressure encountered in being pushed down a borehole.

Bioremediation apparatus 10 is configured at bottom end 16 of casing 12for coupling with an explosive apparatus shown and discussedsubsequently in relation to FIGS. 2–6 as housing a bioremediatableexplosive material. Bioremediation apparatus 10 also has casing threads28 on casing 12 that cooperatively engage correspondingly configuredthreads on the explosive apparatus to effect the intended coupling.

According to teachings of the present invention, microorganisms 30capable of degrading explosive materials are stored in a storage meansfor releasably containing microorganisms. By way of example and notlimitation, such a storage means within the scope of the presentinvention can take the form of a storage chamber 32 having sidewallsdefined by casing 12. As shown in FIGS. 1–5, microorganisms 30 can bepositioned on a ring formed from starch and flour, bran, or anothersimilar nutrient material. The microorganisms 30 can be stored in amoist condition in storage chamber 32 or the microorganisms can also belyophilized or freeze dried. Microorganisms 30 are preferably notmobilized until bioremediation apparatus 10 is actually coupled with anexplosive apparatus which is preferably in the field at the time theseismic charge is to be placed in a borehole or at the time of theintended detonation of the charge in that explosive apparatus. Inaddition to a ring configuration, microorganisms 30 can be positioned incontact with materials such as starch, flour, or bran assuming any otherarrangement.

Microorganisms 30 are mobilized by a liquid 34 stored in a reservoirmeans for releasably containing a liquid. Liquid 34 may be water or anutrient medium that can feed microorganisms 30, but liquid 34 isresistant to freezing at ambient temperatures. By way of example and notlimitation, a reservoir means within the scope of the present inventioncan take the form of a reservoir chamber 36. Reservoir chamber 36 hassidewalls defined by the interior 39 of casing 12 and a top defined bycap 18. Reservoir chamber 36 also includes a liquid passage 38 definedby the interior of a neck 40. Neck 40 is an integral portion of casing12 and has a diameter that tapers radially inwardly from the outerdiameter of the sidewalls of reservoir chamber 36 to a smaller diameter,as observed to best advantage in FIG. 3.

Preferably, storage chamber 32 is positioned below reservoir chamber 36in the anticipated orientation of bioremediation apparatus 10 whencoupled to and installed with explosive apparatus 60. Storage chamber 32is capable of communication with reservoir chamber 36 through liquidpassage 38. Storage chamber 32 is provided with a bioremediation outlet42 that is formed through bottom end 16 of casing 12 and through asleeve member 44 which protrudes from bottom end 16 of casing 12.Accordingly, bioremediation outlet 42 is a portal or opening throughcasing 12 that is in communication with storage chamber 32.

Microorganisms 30 are mobilized by liquid 34 upon the opening of a firstvalve means for delivering liquid 34 from reservoir chamber 36 tostorage chamber 32. By way of example and not limitation, a first valvemeans according to the teachings of the present invention can take theform of a first valve 46 which comprises the interior 39 of neck 40 anda first valve member 48. A tapered end 50 is formed around the perimeterof first valve member 48 corresponding in dimension to the interior 39of neck 40. The cooperation of these structures forms a seal withinliquid passage 38.

When first valve 46, which is shown in FIG. 1, is closed as shown inFIG. 3, first valve 46 is at least partially disposed within reservoirchamber 36. More particularly, when first valve 46 is closed, firstvalve 46 is positioned between reservoir chamber 36 and storage chamber32 and within liquid passage 38 with first valve member 48 defining thebottom of reservoir chamber 36 and the top of storage chamber 32. Theseal formed within liquid passage 38 by first valve 46 thereby retainsliquid 34 in reservoir chamber 36 until first valve 46 is opened.

Once mobilized or activated, microorganisms 30 flow out of storagechamber 32 upon the opening of a second valve means for deliveringmobilized microorganisms to an explosive material in an explosiveapparatus. By way of example and not limitation, a second valve meansaccording to the teachings of the present invention can take the form ofa second valve 52 which includes a second valve member 54 in combinationwith sleeve member 44. Second valve 52 defines the bottom of storagechamber 32 and is the lower end of a valve connector 56 that extendsthrough bioremediation outlet 42 in sleeve member 44. Valve connector 56has an upper end that is connected to first valve member 48.

When second valve 52 is closed as shown in FIG. 3, second valve 52 is atleast partially disposed within storage chamber 32. More particularly,when closed, second valve 52 is positioned at the bottom of storagechamber 32 within bioremediation outlet 42. Second valve member 54 formsa seal with sleeve member 44 in bioremediation outlet 42, thereby toretain mobilized microorganisms 30 in storage chamber 32 until secondvalve 52 is opened. The security of the seal is increased by a valveO-ring 58, which encircles second valve member 54.

Valve connector 56 can best be seen in FIG. 3 to connect first valvemember 48 and second valve member 54. The length of valve connector 56is selected to enable first valve 46 and second valve 52 to open andclose according to a predetermined timing relationship. Thus, firstvalve 46, second valve 52, and valve connector 56 can be configured toeffect the simultaneous opening or actuation of first valve 46 andsecond valve 52. Alternatively, first valve 46, second valve 52, andvalve connector 56 can be designed to actuate one of the valves in adelayed manner, after the other valve has been actuated. It may bedesirable, for example, to allow liquid 34 to contact microorganisms 30by opening first valve 46, and only thereafter to open second valve 52.

When microorganisms 30 and liquid 34 are securely sealed in separatespaces as shown in FIG. 3, bioremediation apparatus 10 can be shippedand stored for long periods without any significant decrease in thebioremediating effectiveness thereof. The configuration ofbioremediation apparatus 10 is designed for easy combination with aconventional explosive, such as a seismic booster.

FIGS. 2 and 3 depict bioremediation apparatus 10 in the process of beingcoupled with an explosive apparatus 60. FIG. 2 is a perspective view,and FIG. 3 is a cross-sectional view taken along section line 3—3 ofFIG. 2. As best illustrated in FIG. 3, first valve member 48 and secondvalve member 54 remain in closed positions when bioremediation apparatus10 is initially threaded into explosive apparatus 60.

Explosive apparatus 60 comprises a shell 62 having an open end 64 and anexplosive material 66 housed within shell 62. On an extension 68 ofshell 62 not surrounding explosive material 66, the interior of shell 62is provided with shell threads 70 near open end 64. These cooperate withcorrespondingly configured casing threads 28 on bioremediation apparatus10. Shell 62 can be formed from distinct components or as an integralstructure as shown.

The combination of casing threads 28 and shell threads 70 together serveas an example of a coupling means for coupling a bioremediationapparatus according to the teachings of the present invention with anexplosive apparatus, such as explosive apparatus 60. In the embodimentillustrated, the function of such a coupling means is performed by anextension of casing 12 of bioremediation apparatus 10 and extension 68of shell 62 of explosive apparatus 60. Alternatively configuredstructures can, however, perform the function of such a coupling means.

For example, a wedge fit can be effected between bioremediationapparatus 10 and explosive apparatus 60 using respective angled male andfemale parts attached, respectively, to each. While the coupling meansis primarily a mechanism to join bioremediation apparatus 10 andexplosive apparatus 60, it is within the teachings of the presentinvention to provide structures that prevent bioremediation apparatus 10and explosive apparatus 60 from being unintentionally separated, therebyperforming the function of a locking means for securing bioremediationapparatus 10 and explosive apparatus 60 against the disengagement of thecoupling together thereof.

Explosive apparatus 60 further comprises a capwell 72 positioned in openend 64 to receive detonators 74. Detonators 74 are in turn electricallyconnected by wires 76 to the exterior of shell 62 through wire accessopenings 78 shown in FIG. 2. A bioremediation portal 80 formed throughcapwell 72 communicates with explosive material 66 to afford access bymobilized microorganisms 30 from bioremediation outlet 42 to explosivematerial 66. A portal member 82 extends upwardly as shown in FIG. 3 fromthe center of capwell 72, encircling and defining on the interiorthereof bioremediation portal 80. A portal O-ring 84 encircles portalmember 82 to provide a fluid seal between sleeve member 44 and portalmember 82 when explosive bioremediation apparatus 10 is coupled withexplosive apparatus 60.

FIGS. 4 and 5 depict bioremediation apparatus 10 immediately afterbecoming completely coupled with explosive apparatus 60. FIG. 4 is aperspective view, and FIG. 5 is a cross-sectional view taken alongsection line 5—5 of FIG. 4. FIG. 5 illustrates to best advantage that asa result of the coupling of bioremediation apparatus 10 with explosiveapparatus 60, first valve 46 and second valve 52 have been opened.Liquid 34 is shown being delivered by gravity from reservoir chamber 36through liquid passage 38 to storage chamber 32.

As bioremediation apparatus 10 is being coupled with explosive apparatus60, sleeve member 44 and portal member 82 advance toward each otheruntil portal member 82 is positioned within sleeve member 44. A fluidseal is formed between sleeve member 44 and portal member 82 by portalO-ring 84. The advancement of bioremediation apparatus 10 brings portalmember 82 into abutment against second valve member 54. As valveconnector 56 effects a rigid interconnected relationship between secondvalve member 54 and first valve member 48, further advancement ofbioremediation apparatus 10 into and toward explosive apparatus 60forces second valve member 54 out of bioremediation outlet 42 and forcesfirst valve member 48 out of liquid passage 38.

Sleeve member 44, second valve member 54, and portal member 82 can haveany lengths that enable first valve 46 and second valve 52 to be opened.In the embodiment shown in FIGS. 5 and 6, portal member 82 and secondvalve member 54 each have a length that is less than the length ofsleeve member 44, and the length of portal member 82 is approximatelyequal to or greater than the length of second valve member 54.

Forcing second valve member 54 and first valve member 48 upward withincasing 12 opens a flow path that permits mobilized microorganisms 30 tocontact explosive material 66 through bioremediation portal 80 as shownin FIG. 6.

First valve member 48, second valve member 54, and valve connector 56form a divider which is preferably formed at least partially from alightweight material such as polyethylene. The divider in this mannerpreferably has a lower density than water. This enables the divider tofloat to the top of the suspension of microorganisms as shown in FIG. 6after the liquid has flowed into contact with explosive material 66.

The divider formed by first valve member 48, second valve member 54, andvalve connector 56 is an example of a divider means for releasingmicroorganisms to an explosive material according to the teachings ofthe present invention. In an alternative embodiment, the divider meanscan take the form of a valve means for delivering the mobilizedmicroorganisms from a storage means to an explosive material in astorage chamber of an explosive apparatus. The microorganisms are in amoist condition or are in a sufficient quantity of liquid to becharacterized as a suspension or dispersion. Such an alternativeembodiment accordingly utilizes but a single chamber and a single valve.

Portal member 82 is an example of an actuation means for initiatingcontact between mobilized microorganisms and an explosive material byopening first valve 46 and second valve 52. In an alternativeembodiment, portal member 82 has a length greater than sleeve member 44,thereby rendering unnecessary any second valve member extending withinsleeve member 44 to align portal member 82 for contact with the secondvalve member. In an additional alternative embodiment, second valve 52is configured similarly to first valve 46. In this additionalalternative embodiment, second valve 52 has a valve member withinbioremediation outlet 42 that does not extend downward, and there is nosleeve member to provide alignment for portal member 82 in contactingthe valve member to actuate second valve 52. Any structure capable ofinitiating access by mobilized microorganisms 30 to explosive material66 is within the scope of the actuation means of the present invention.

The actuation means and the coupling means are taken together exemplaryof a contact means for initiating and maintaining contact betweenmobilized microorganisms and an explosive material.

The coupling of bioremediation apparatus 10 with explosive apparatus 60forms a system for in situ bioremediating of an explosive material. Thesystem can be lowered into or driven down a borehole by contacting cap18 with a tamping pole. Additionally, an anchor member 90 shown only inFIG. 6 can be positioned about casing 12 to maintain the system in theupright position illustrated during installation of the system at thebottom of a borehole. Anchor member 90 is preferably a disccircumferentially encircling casing 12 and extending perpendicularlyoutwardly therefrom. The longitudinal position of anchor member 90 alongthe length of casing 12 is maintained as shown in FIG. 6 by the increasein the outer diameter of casing 12 above anchor member 90.

The failure of installed explosives to detonate is primarily caused bythe forces experienced during positioning of the system in the bottom ofa borehole. In the process, wires 76 are often broken or disconnectedfrom detonators 74, so that detonation cannot occur. When this happens,the digestion of explosive material 66 by microorganisms 30 will proceedin due course. Eventually, explosive material 66 will be reduced tononexplosive and non-harmful materials that are neither detonatable byany activities in the vicinity, nor are an environmental contaminant.

Over time, by exposing an undetonated charge to the microorganisms, theentirety of the explosive material of the charge is reduced to asubstance that cannot be detonated. In the illustrated embodiments ofthe present invention, the digestive activity of microorganisms 30disarms explosive material 66 by first attacking the area around thecapwell end of the explosive apparatus. This is where detonation isactually initiated. There is, however, no overall detrimental effect onthe ability of an explosive charge to be detonated immediately afterbeing initially contacted by bioremediating microorganisms. The initialactivity of the microorganisms in the vicinity of the capwell canprevent accidental detonation of the explosive charge which can becaused, for example, by digging in the area of the explosive chargeafter the explosive charge is positioned in a borehole.

The time period required for the microorganisms to first disable anexplosive and then to fully remediate a given quantity of intermediatechemical materials depends on the amount and type of explosive materialused, as well as the composition of microorganism consortium usedtherewith. Depending on design and relative concentrations of theexplosive, the time required can be days, weeks, months, or years.

FIGS. 7 and 8 depict a second embodiment of a system for in situbioremediating of an explosive according to teachings of the presentinvention. The system shown there comprises a bioremediation apparatus100 and an explosive apparatus 110. Components shown in FIGS. 7 and 8that are identical to the components shown in FIGS. 1–6 are identifiedwith the same reference characters as are the corresponding componentsin FIGS. 1–6.

Bioremediation apparatus 100 has a cap 18, a spacer 112, and an anchormember 90 that encircles top end 14 of casing 12. Cap 18 and a spacer112 are configured to maintain anchor member 90 on a nib 114. Cap 18 hascap threads 24 which cooperate with end threads 26 around top end 14 ofcasing 12 to seal top end 14 of casing 12. Spacer 112 is positionedbetween cap 18 and anchor member 90. Spacer 112 has a bottom portion notshown in the figures that is positioned within the top end of anchormember 90. When the system of FIGS. 7 and 8 is pushed down a boreholewith a tamping pole, anchor member 90 cannot be dislodged from nib 114,since anchor member 90 abuts spacer 112, and cap 18 retains spacer 112in position.

Liquid 34 is contained in reservoir chamber 36 and is released tocontact microorganisms 30 in storage chamber 32 when a first valve 116in liquid passage 38 is opened. First valve 116 comprises the interiorof neck 40 and a first valve member 118. First valve member 118 has alip end 120 around the perimeter of first valve member 118. Lip end 120is tapered to correspond to the dimensions of the interior of neck 40and is flexible, thereby to form a fluid seal with liquid passage 38.

A second valve 122 comprises a second valve member 124 and a lip sealmember 126. Second valve member 124 is the tapered bottom end of a valveconnector 128. Lip seal member 126 extends from sleeve member 44 intobioremediation outlet 42 to form a fluid seal with second valve member124.

First valve member 118 is integrally formed with valve connector 128,and valve connector 128 is integral with second valve member 124.Accordingly, first valve member 118, valve connector 128, and secondvalve member 124 together form an integral divider. In the firstembodiment shown in FIGS. 1–6, the first valve means is also connectedto the second valve means, as first valve member 48 and second valvemember 54 are connected by valve connector 56. Thus, both in the firstembodiment of FIGS. 1–6 and in the second embodiment of FIGS. 1–8, atleast a portion or a component of each valve means is connected to atleast a portion or a component of the other valve means.

The coupling means for coupling a bioremediation apparatus with anexplosive apparatus, such as the combination of casing threads 28 andshell threads 70 as shown in FIGS. 1–8, may further comprise a means forindicating the position of the valves. In the preferred embodiment,casing 12 preferably has a bump not shown in the figures that causes aclicking noise when portal member 82 contacts the second valve memberafter casing threads 28 and shell threads 70 are advanced over eachother. The clicking noise informs a user that the bioremediationapparatus and the explosive apparatus are coupled.

In yet additional alternative embodiments of an apparatus similar to theapparatus depicted in FIGS. 1–6 or in FIGS. 7–8, the first or secondvalve means can be electronically controlled to remain closed untilelectronically activated. For instance, a battery can provideelectricity to retain a valve in a closed position such that the valveopens when the battery is dead. Accordingly, the microorganisms are notmobilized until after a time period equal to or exceeding the life spanof the battery. In such an embodiment it is unnecessary to couple abioremediation apparatus and explosive apparatus together to mobilizethe microorganisms so the microorganisms and explosives can be containedin a single housing.

An example of an embodiment that utilizes an electrical mechanism isshown in FIG. 9 which is similar to the embodiment shown in FIGS. 7–8.Bioremediation apparatus 140 shown in FIG. 9 has microorganisms 30 whichare not in the doughnut configuration but are added as a block. A flange142 extends from valve connector 56 and is above a piston 144. Piston144 extends within a spring 146 from a solenoid 148. Solenoid 148 iselectrically connected to a battery 150. Battery 150 applies power tothe coil of solenoid 148 which pulls piston 144 into the coil ofsolenoid 148 and retains piston 144 against the force of spring 146 aslong as power is being supplied to the coil of solenoid 148. When thebattery is dead then piston 144 and spring 146 are released and pushagainst flange 142 which causes first valve member 48 and second valvemember 54 to be pushed upward, thereby opening first valve 46 and secondvalve 52. In this embodiment, it is not necessary for sleeve member 44,second valve member 124 or portal member 82 to have lengths that enablethe valve members to be opened.

Other embodiments utilize mechanisms which are primarily chemical innature such as a barrier formed from a material, which is water solubleor slowly self-effacing in the presence of water, an aqueous solution ormicroorganisms. Thus, for example, self effacing barriers whicheventually degrade and release the microorganisms can perform thefunctions of either or both first or second valve means according to thepresent invention. Examples of materials that can be utilized asself-effacing valve members include gelatin, alginate, starch, andacrylamide. Careful structural and material design of such barriers canproduce relatively precisely timed releases. Alternatively,microorganisms encapsulated in a material such as gelatin or alginatemay be releasably contained in a storage means of the present inventionfor eventual contact with an explosive material.

FIGS. 10–16 depict embodiments of the present invention whereinmicroorganisms are intermixed in the explosive or are disposed againstan exterior surface of the explosive material. The microorganismsdepicted in FIGS. 10–16 are disposed in sufficient proximity to saidquantity of explosive material that the microorganisms can initiatebioremediation of the explosive material when the microorganisms aremobile. The explosive apparatus shown in FIGS. 10–16 do not require thecoupling of a distinct bioremediation apparatus with a correspondingexplosive apparatus.

The microorganisms intermixed in the explosive material are generally inaggregations or clusters such as pellets as shown in FIG. 10, capsulesas shown in FIGS. 11–12, or shards as shown in FIG. 13. The embodimentsdepicted in FIGS. 14–16 provide examples of microorganisms disposedagainst an exterior surface of the explosive material. FIG. 14 shows apowder of microorganisms dispersed on the top surface of the explosivematerial. FIG. 15 depicts microorganisms poured into a column within theexplosive material. FIG. 16 depicts a cluster of microorganismspositioned within the shell that contains the microorganisms. Inaddition to the clusters or aggregations disclosed in FIGS. 10–16, themicroorganisms can be positioned in any form even as individualmicroorganisms.

FIG. 10 illustrates an explosive apparatus 200 configured with anoptional cap 202 and access openings 204 for wires 76. As in theembodiments depicted in FIGS. 1–9, explosive apparatus 200 has a capwell72 with detonators 74. Explosive apparatus 200 further comprises a shell206 containing an explosive material 208 and pellets 210 ofmicroorganisms dispersed throughout explosive material 208. Shell 206preferably enables water to flow through shell 206 to contact theexplosive material 208 or at least into contact with the microorganismsin pellets 210 at the exterior surfaces of explosive material 208. Shell206 may for example have an open end wherein water can flow, have holesor be water permeable to enable water to enter into the pores ofexplosive material 208.

Pellets 210 are dispersed as needed. For example, pellets 210 can berandomly dispersed, as shown, or concentrated as needed to deactivatethe explosive charge. Pellets 210 are preferably positioned tofacilitate desensitization of the explosive apparatus by beingconcentrated within explosive material 208 around detonators 74.

Pellets 210 can be positioned within explosive material 208 by anymethod and in any desired concentration. Control of the concentrationand dispersion of pellets 210 in the explosive material 208 is maximizedby adding pellets 210 to explosive material 208 when explosive material208 is in a liquid state. Explosive material 208 is in a liquid statewhen being formed into a desired configuration by pouring the explosivematerial into a mold or directly into shell 206. The forming temperatureof the explosive material is around 100° C. which is generally lethal tothe microorganisms. Accordingly, the exposure time of microorganisms inpellets 210 to lethal temperatures is preferably minimized by addingpellets 210 to explosive material 208 while explosive material 208 isbeing formed or cast into a desired shape. Pellets 210 can also bepressed into explosive material 208 when explosive material 208 is solidor semi-solid at the time that the charge is manufactured.

The microorganisms or pellets 210 containing the microorganisms arepreferably heat resistant to increase the survivability of themicroorganisms when added to explosive material 208. There are severalmethods, which can be utilized alone or in combination, for obtainingheat resistant microorganisms or pellets.

One method for obtaining heat resistant microorganisms involveslyophilizing the microorganisms before the microorganisms are added tothe hot explosive material. The microorganisms can be dehydrated byallowing the water to evaporate or preferably by freeze drying themicroorganisms. Freeze drying the microorganisms dramatically reducesthe mortality of the microorganisms due to thermal stress from exposureto the molten explosive material during the pouring process. It isspeculated that freeze dried microorganisms are less susceptible to thelethal temperature effects than a microorganisms in a moist environmentbecause the water content in the moist microorganism provides betterheat transfer to the vital and temperature sensitive internalstructures. The water removed from the freeze dried microorganisms isreplaced at a later time in sufficient quantity to activate and mobilizethe microorganisms.

The survivability of the microorganisms to thermal stress is alsoincreased by increasing the thickness of the pellets 210. Increasing thethickness of pellets 210 decreases the rate of heat transfer to theinterior of pellets 210, thereby protecting the microorganisms in theinterior to the extent that the residence time of the microorganisms inthe hot melt is not excessive. When the exterior microorganisms aredestroyed they act as a thermal insulator for the microorganisms withinthe interior. Suitable pellets generally have an average diameter ofabout 3 mm.

Another method for reducing the mortality of microorganisms due tothermal stress is achieved by adding the microorganisms and explosivematerial into a mold in thin layers. By adding the microorganisms andexplosive material incrementally in thin layers the layers can quicklycool thereby minimizing the exposure time of the microorganisms to thehot melt.

The heat resistance can also be increased by slowly raising the growthtemperature of the microorganisms. By slowly raising the temperature ofthe environment of the microorganisms over a period of time during theirgrowth, the high temperature tolerance of the microorganisms issignificantly increased. Microorganisms can be utilized which have beenadaptively developed or which have also been genetically developed. Themicroorganisms are preferably developed to have a very highsurvivability rate even when exposed to temperatures as high as about100° C. Even microorganisms which have not been adequately developed tosurvive exposure to temperatures as high as about 100° C. are veryuseful since the temperature decreases as it is transferred into thepellet, yielding a greater interior portion that survive compared to apellet utilizing unconditioned microorganisms. Accordingly, allmicroorganisms that have been developed for high temperature toleranceare useful and yield a higher survivability rate.

Additionally, pellets 210 can also be formed from mixtures ofmicroorganisms and thermal protection additives that increase the heatresistance of the microorganisms. Examples of additives which have beenfound to increase the survivability of microorganisms when thermallystressed include dry milk and bentonite clay. These viability enhancersalso can be used as binders as they tend to bind the constituents in thepellet together and can also be a nutrient source for themicroorganisms. Insulative aggregates with no binding capability or thatare not nutrient sources can also be used to thermally protect themicroorganisms.

The pellets can be formed by compressing the microorganisms togetheralong with any other constituent materials such as nutrients, bindersand insulative materials. As previously set forth, the same componentcan act as a nutrient, binder or an insulative material. Nutrients aregenerally necessary even when the microorganisms are lyophilized sincethey provide the microorganisms with all of the materials needed for themicroorganisms to fully grow and multiply. The explosives generallyprovide carbon and nitrogen while the nutrients generally providephosphate and other chemicals. Any suitable nutrient can be utilized;however, depending on the type of nutrient utilized and the availabilityof the nutrient the growth rate can be influenced. In addition to thenutrients previously discussed such as starch, flour, bran, and milk;other suitable nutrients include milk sugar and minimal medium glycerol.Many of these nutrients can also act as stabilizers, such as starch,flour, bran, milk, glycerol in addition to phosphate buffered saline.

It is not always necessary to add a component that acts only as a bindersince many nutrients can be utilized as a binder which is then convertedinto nutrients when contacted by a sufficient quantity of water tosolubilize the binder. In addition to the binders previously mentioned,any suitable binder can be utilized. The binder is preferably an organicbinder. A product sold as Diatab is a particularly useful binder ortablet base. Other materials that can be utilized as a binder includeacrylamide, alginic acid or alginate, ethylcellulose, guar gum andgelatin.

Pellets 210 can also be encapsulated in a capsule 212 as shown in FIG.11. The microorganisms in capsule 212 can be freeze dried, then formedinto a pellet and encapsulated or the capsule can be formed byencapsulating moist microorganisms or a suspension of microorganism bypouring the suspension into a capsule and then drying or freeze dryingthe capsule. Any suitable materials for forming capsule 212 can beutilized. Examples of suitable materials include gelatin, starch,alginate and acrylamide.

While it is preferred to form the explosive by placing pellets ofdehydrated microorganisms into molten explosive material since themolten explosive material is easily and relatively quickly molded into adesired shape, it can decrease the viability of the microorganisms.Accordingly, it is also desirable to form pellets 210 from moistmicroorganisms. Depending on the amount of liquid present, it may benecessary to introduce the microorganisms into explosive material 208 asa suspension 214 of microorganisms shown in FIG. 12 that is contained ina capsule 212.

Microorganisms which are merely moist can also be encapsulated or can beintroduced as shards 216 of a moist nutrient wafer containingmicroorganisms as shown in FIG. 13. Shards 216, which are fragments orflakes, can also be a nutrient wafer containing lyophilizedmicroorganisms.

Clusters or aggregations of moist microorganisms in configurations suchas pellets, capsules, shards, flakes and the like are preferably blendedinto explosive material 208 and then pressed into a mold. Moistmicroorganism clusters can also be pressed into explosive material 208.When the microorganisms are in a moist state but are blocked fromcontact with the explosive material or are not sufficiently mobile,nutrients provide a minimal food source until the microorganisms canmetabolize the explosive material. After the mixture is shaped into anexplosive charge and the microorganisms are sufficiently moist, it willbioremediate automatically within a predetermined time followingmanufacture.

An explosive apparatus is often left underground for periods of time upto six months and even up to a year. Accordingly, an explosive apparatusis preferably explodable for up to about six months and more preferablyfor up to about a year.

As previously set forth, the microorganisms can be concentrated arounddetonators 74 to desensitize the explosive since detonators 74 aretypically more sensitive to impact and friction than explosive material208. The time required to desensitize explosive apparatus 200 bydisabling explosive material 208 around detonators 74 is dependent onmany variables in addition to the distribution of the microorganisms,the growth rate of the types of microorganisms utilized, the ratio ofmicroorganisms to explosives, the availability of particular nutrients,the types of microorganisms and explosives utilized and other physicalcondition such as pH, water availability and temperature. These samevariables generally determine the time required to reduce explosivematerial 208 to a residual or negligible amount and the time required toentirely reduce explosive material 208 to a nonhazardous and preferablynonharmful material. Some of these additional variables include theamount of surface area exposed to the microorganisms, the mobility ofthe microorganisms, and the porosity of the explosive materials.Accordingly, the bioremediation rate can be designed as needed.

The porosity of the explosive materials is an example of a mobilizationmeans for mobilizing the microorganisms to contact explosive material208. The porosity of the explosive material 30 enables the mobilizedmicroorganisms to move within explosive material 208 and continuebioremediating explosive material 208. The porosity also enables waterto enter into the pores and come into contact with the microorganismsand mobilize the microorganisms. A surfactant in explosive material 208is another example of a mobilization means. Surfactants facilitatewetting of the crystals in explosive material 208 which enhances themobility of the microorganisms and the accessibility of the crystals tothe microorganisms.

Explosive apparatus 200 can be immersed in water before being placed ina borehole to allow water to pass through shell 206 and enter into thepores to mobilize the microorganisms or clusters thereof intermixed inexplosive material 208. Explosive apparatus 200 is preferably exposed toa vacuum before being dipped in water. It is generally not necessary toimmerse explosive apparatus 200 in water as groundwater is almost alwayspresent in the borehole. Additionally, water can also be poured into theborehole as needed. Water around or in contact with explosive apparatus200 are additional examples of mobilization means for mobilizing themicroorganisms.

Since groundwater is almost always in a borehole, it is generallydesirable to design the explosive apparatus to utilize the groundwater.Accordingly, the porosity is preferably conducive to optimal capillaryaction through a network of microchannels. The network of microchannelsor pores is sufficiently interconnected to provide optimal accessibilityto the microorganisms by water and to provide optimal mobility to themobilized microorganisms. The porosity is also designed to provideoptimal surface area for the microorganisms to bioremediate. Theporosity is balanced against the amount of explosive material that ispreferably present and any necessary amount of mechanical strength forwithstanding crushing and other forces experienced while beingpositioned in the borehole. The porosity can also be heterogenousthroughout explosive material 208 such that the area around detonators74 is more porous compared to other sections to expose more surfacearea.

The embodiments depicted in FIG. 10–13 are dependent primarily on theporosity of explosive material 208 to provide access to themicroorganisms and to provide mobilization pathways for the mobilizedmicroorganisms. FIGS. 14–15 depict embodiments of the present inventionthat do not rely primarily on the porosity of explosive material 208.

FIG. 14 depicts microorganisms deposited as granules 218 on top ofexplosive material 208. Accordingly, as water passes through shell 206the initial bioremediation activity of all of the microorganisms isconcentrated at the portion of explosive material around detonators 74.

FIG. 15 depicts a chamber 220 centrally and longitudinally locatedwithin explosive material 208 that contains a suspension 222 ofmicroorganisms. Microorganisms can also be positioned in chamber 220which are merely moist or have been lyophilized. This configurationenables the mobilized microorganisms to bioremediate explosive material208 from within a particular location in explosive material. Theposition of chamber 220 provides for controlled bioremediation ofexplosive material 208 around detonators 74.

FIG. 16 depicts another embodiment wherein shell 62 contains clumps 224of microorganisms. Shell 62 is preferably formed from a material that isnot only water permeable but also sufficiently water soluble to releasethe microorganisms contained in the shell. Examples of suitablematerials include but are not limited to paper and polyvinyl alcohol.The microorganisms can then bioremediate explosive material 208 bybeginning at the exterior of explosive material 208.

Yet another method of bioremediating explosives involves installing anexplosive charge in a detonation site, such as a borehole, and thenpositioning microorganisms around the explosive charge by depositingmicroorganisms directly on the explosive charge and the detonation site.Similarly, a solution of microorganisms can be deposited at a detonationsite. Then the explosive charge is placed in the suspension ofmicroorganisms. Additionally, an explosive apparatus can be sprayed withor soaked in a suspension of microorganisms before being installed at agiven detonation site, preferably while being exposed to a vacuum.

Experiments were conducted to study the process of remediating explosivematerials according to the teachings of the present invention. To do so,a microorganism consortium was derived from soil and water samplesobtained on the property of an established explosive manufacturerlocated at 8305 South Highway 6, Spanish Fork, Utah 84660 U.S.A. Themicroorganism consortium in the form of a suspension was combined withvarious types of explosive materials, either in solid form or in anaqueous suspension, and the results were observed and documented. Theresults of several of these tests are set forth below as examples.

EXAMPLE 1

Quantities of the explosive materials TNT and PETN in water werecombined with the suspension of the microorganism consortium. Theresulting mixture initially included 47.23 parts per million (ppm) ofPETN and 40.63 ppm of TNT. The mixture was divided among containers thatwere stored in aerobic conditions at ambient temperature for varioustime periods. Table 1 below indicates the explosive analysis of thesesamples after each designated time interval. The explosive materialswere substantially degraded after a period of five weeks.

TABLE 1 Aerobic Bioremediation of TNT and PETN Explosive Analysis AfterAnalysis After Material Initial Analysis 3 Days 5 Weeks PETN 47.23 ppm40.94 ppm 7.25 ppm TNT 40.63 ppm  5.32 ppm 0.62 ppm

EXAMPLE 2

The mixture prepared in Example 1 was stored in anaerobic conditions atambient temperature and observed. The results were determined by HPLCanalysis in ppm and averaged. Table 2 below sets forth the resultsobtained. As can be seen by comparing the results in Table 2 with theresults in Table 1, the explosive materials tested remediated morerapidly under anaerobic conditions than under aerobic conditions.

TABLE 2 Anaerobic Bioremediation of PETN and TNT Explosive InitialAnalysis after Analysis after Analysis after 5 Material Analysis 3 Days1 Week Weeks PETN 47.23 ppm 28.31 ppm 24.46 ppm 0.82 ppm TNT 40.63 ppm 0.31 ppm avg.  0.31 ppm avg. None

EXAMPLE 3

Discs of the explosive material Pentolite having a diameter of a pencilwere split in two. When the discs were split each weighed about 0.1gram. The discs were placed either in water as a control or in 6 ml to 8ml of a suspension of the microorganism consortium. After a specificamount of time in aerobic conditions, the discs were dried and weighedor analyzed by HPLC. The liquid portions were analyzed by HPLC. The netremediated weight loss in the explosive material was determined bysubtracting the control weight loss as a percentage from the weight lossas a percentage in each remediated explosive. The explosive loss bydegradation is listed in Table 3 for each of the samples. The samples inB and C were tested for longer periods of time than the sample in A. Theresults of the testing of samples B and C show that significantbioremediation did not occur beyond the level achieved in sample A. Thiswas most likely due to insufficient quantities of nutrients in samples Band C as the bioremediation activity probably ceased when the nutrientswere consumed.

TABLE 3 Aerobic Bioremediation of Pentolite Final dry weight plus weightof Sample Sample or explosive in Net Remediated No. Test Time InitialWeight liquid portion. Weight Loss A Control  22 days 0.1355 g 0.1266 g= 6.57% 6.97% Net Loss loss Test  22 days 0.0981 g 0.0848 g = 13.54%loss B Control  88 days 0.0578 g 0.0557 g = 3.63% 5.52% Net lossExplosive Loss Test  88 days 0.0743 g 0.0675 g = 9.15% loss C Control173 days 0.1236 g 0.1236 g = no 6.78% Net loss Explosive Loss Test 173days 0.0737 g 0.0687 g = 6.78% loss

EXAMPLE 4

Experiments were conducted to compare remediation rates under aerobicand anaerobic conditions. Separate 5 gram samples of PETN/TNT Pentolitein a ratio of 60:40 were analyzed and placed in 100 ml to 300 mlsuspension of a microorganism consortium. One was subjected to aerobicconditions; the other was subjected to anaerobic conditions. Aftervarious periods of time the samples were removed, air dried, and weighedto determine the amount of explosive material that had not degraded. Theweight of the remaining explosive material was subtracted from theoriginal weight to determine the weight of the explosive material lostdue to bioremediation. The results are listed in Table 4 below. Theresults indicate that an insufficient amount of microorganisms wereutilized or that the amount of nutrient was insufficient, particularlyin light of the results obtained in the other examples.

TABLE 4 Aerobic and Anaerobic Bioremediation of Pentolite Condition:Percent Wt Percent Aerobic or Original Loss at Wt Loss at AnaerobicWeight Time Time listed Time Time Listed Aerobic  5.015 g 66 days 3.21%163 days 5.43% Anaerobic 6.9027 g — — 179 days 3.10%

EXAMPLE 5

Also investigated was the remediation according to the present inventionof low levels of explosive materials in water. The explosive materialsRDX and PETN were mixed with the water, combined with a suspension of amicroorganism consortium, and then stored. The samples were tested byHPLC for explosive content initially and after 2 weeks. As shown inTable 5 below the bioremediation was nearly complete after two weeks.

TABLE 5 Bioremediation of Suspension of RDX and PETN Explosive MaterialInitial Analysis Analysis after 2 weeks RDX  6.6 ppm Not detected PETN25.0 ppm Less than 0.5 ppm

EXAMPLE 6

The remediation according to the present invention of soil contaminatedwith an explosive material was also investigated. Soil contaminated withthe explosive material PETN was mixed with a suspension of amicroorganism consortium and stored at ambient temperature. Samples wereanalyzed initially, after 44 days, and finally after 125 days. The PETNcontent in the soil dropped from 1659 ppm to 551 ppm. The results areset forth in Table 6 below.

TABLE 6 Bioremediation of Soil Contaminated with PETN Initial AnalysisAnalysis after 44 Days Analysis after 125 Days 1659.2 ppm 1193.2 ppm551.8 ppm

EXAMPLE 7

In order to determine the effect of temperature on the growth ofmicroorganism samples, the natural high temperature tolerances of themicroorganism consortium were evaluated. The microorganism cultures wereadapted to higher temperatures by slowly raising the growth temperature.By raising the temperature, the upper and lower limits of growth wereboth shifted upwards.

Two separate microbial growth stages were evaluated: the log phase,wherein the microorganisms experience logarithmic growth, and thestationary phase, wherein the microorganisms reach maximum growth.Microorganism cultures that enter the stationary phase late in theirgrowth cycle induce the expression of genes which protect themicroorganisms from various environmental stresses.

Four separate microorganism cultures were established. One culture,referred to as “30° C./Log Phase Culture,” was comprised of new inocula,experiencing logarithmic growth, in fresh minimal medium, with TNTextract as the sole nitrogen source, and grown at 30° C. for three days.A second culture, referred to as “30° C./Stationary Phase Culture,” wascomprised of microorganisms that had reached maximum growth, in minimalmedium, with TNT extract as the sole nitrogen source, previously grownat room temperature for several weeks, and additionally grown at 30° C.for three days. The third culture, referred to as “37° C./Log PhaseCulture,” was comprised of new inocula, experiencing logarithmic growth,in fresh minimal medium, with TNT extract as the sole nitrogen source,and grown at 37° C. for three days. The final culture, referred to as“37° C./Stationary Phase Culture,” was comprised of microorganisms thathad reached maximum growth, in minimal medium, with TNT extract as thesole nitrogen source, previously grown at room temperature for severalweeks, and additionally grown at 37° C. for three days.

Samples of the four different microorganism cultures were subjected totemperatures ranging from 30° C. to 97° C. for twenty minutes. A smallsample of each heated culture and a non-heated control culture werespread-plated on both nutrient agar plates, and minimal medium with 10%glycerol plates. The plates were incubated overnight at 30° C.

The microbial growth was evaluated according to the number of colonyforming units of the plate or the visualization of distinct colonies.The results of this evaluation are illustrated in Table 7 below.Microbial growth covering the entire plate with few, if any, singlecolonies, was referred to as “total.” Microbial growth greater than 1000clearly defined colonies per plate, or too numerous to count, wasreferred to as “>1000”. If the density of the sample was only slightlyless than the density of the previous sample, an asterisk “*” appearsafter the notation. At the lower density levels, the colonies weredistinguishable as comprising at least bacteria, “B” orfungus/filamentous bacteria, “F.” The number preceding “B” or “F”corresponds to the number of distinct colonies.

TABLE 7 Temperature tolerance of microorganism consortium. 30° C./LogPhase 30° C./Stationary 37° C./Log Phase 37° C./Stationary Temp. ° C.Culture Phase Culture Culture Phase Culture Control Total >1000Total >1000 30° C. Total >1000 Total >1000 37° C. Total >1000Total >1000 42° C. Total  >1000* Total >1000 47° C. Total*  >1000*Total* >1000 52° C. 130 B 180 F Total* 7 F 57° C. 0 colonies 0 colonies0 colonies 0 colonies 62° C. 0 colonies 0 colonies 0 colonies 0 colonies67° C. 0 colonies 0 colonies 2 colonies 0 colonies Control Total >1000Total >1000 72° C. 0 colonies 0 colonies 0 colonies 0 colonies 77° C. 0colonies 0 colonies 0 colonies 0 colonies 82° C. 2 colonies 0 colonies 1colony 0 colonies 87° C. 0 colonies 0 colonies 0 colonies 0 colonies 92°C. 0 colonies 0 colonies 0 colonies 0 colonies 97° C. 0 colonies 0colonies 0 colonies 0 colonies Control Total >1000 Total >1000

The log phase cultures appeared predominantly to contain a single colonytype of microorganism. The stationary phase cultures contained a singlemicroorganism colony type and an organism that appeared to be a fungusor a filamentous bacterium.

None of the heated culture samples exhibited significant growth beyond57° C. The difference in the growth phase of the cultures, i.e., logphase versus stationary phase, did not result in a significantdifference in growth. However, the 37° C./Log Phase Culture did appearto exhibit some growth advantage. Note that at 52°, the 37° C./Log PhaseCulture still had microbial growth covering the entire plate, whereasthe growth of the other samples had been reduced to countablequantities.

In addition, the 37° C./Stationary phase Culture and 37° C./Log phaseCulture samples exhibited a growth advantage over the 30° C./StationaryPhase Culture and 30° C./Log Phase Culture which is commensurate withthe differential initial growth temperature of these samples. That is,because microorganism cultures can be adapted to higher temperatureswithin limits by slowly raising or lowering the growth temperature, byraising the temperature, the upper and lower limits of growth are bothshifted upwards. Thus the 37° samples were amenable to more substantialgrowth at higher temperatures than the 30° samples.

Along these lines, a new culture of the 37° C./Log Phase Culture wasestablished using minimal medium with TNT. A sample of this culture wasplaced in a water bath wherein the temperature was raised 1° C. everytwo days. Significant growth was exhibited as high as 41° C.

EXAMPLE 8

In order to assess the survival characteristics of the microorganismculture during cooling of the explosive charge, the following simulatedcasting experiment was performed using the 37° C./Log Phase Culture.Small samples of this culture were placed in tubes in water baths at 95°C. and 80° C. These water baths were programmed to drop 1° C. everyminute based on a reasonable approximation of the rate of coolingexperienced by the charge. At five minute intervals, small samples wereremoved from the tubes in the water baths and plated on nutrient agarplates. These plates were incubated at 30° C. overnight and checked at12 and 36 hours for microorganism colonies. After 36 hours the growth onthe plates was evaluated. A non-heated sample was included as thecontrol. The results of this study are illustrated in Table 8 below.

The results of this study indicate that the samples from the 80° C.water bath had a better survival rate than the samples from the 95° C.water bath.

TABLE 8 Temperature tolerance of microorganism consortium in simulatedcasting. Temperature Time 95° C. Bath 80° C. Bath Control  0 min TotalTotal 90° C.  5 min 0 colonies NA 85° C. 10 min 1 colony NA 80° C. 15min 0 colonies NA 75° C. 20 min/5 min 0 colonies 1 colony 70° C. 25min/10 min 1 colony 2 colonies 65° C. 30 min/15 min 1 colony 1 colony60° C. 35 min/20 min 1 colony 3 colonies 55° C. 40 min/25 min 0 colonies1 colony 50° C. 45 min/30 min 0 colonies 4 colonies 45° C. NA/35 min NA3 colonies 40° C. NA/40 min NA 1 colony 35° C. NA/45 min NA 5 colonies

EXAMPLE 9

The purpose of the following evaluation was to demonstrate that anygrowth on TNT was greater than that which might be expected from lowlevel contamination by nitrogen from other sources. In order to evaluatethe growth characteristics of the microorganism culture with respect tothe nitrogen supply, the following experiment was performed underaerobic conditions.

A sample of the 37° C./Log Phase Culture was placed in each of threefresh media formulations. The first contained mineral salts definedmedium (MMO) and ammonia as the nitrogen source. The second containedMMO and TNT as the nitrogen source. The third contained only MMO and noadded nitrogen. The cultures were then grown and shaken in an incubatorat 37° C.

Growth was measured by evaluating the optical density of the culture.Samples removed from each culture were placed in a spectrophotometer andthe optical density was measured at a wavelength of 425 nanometers, awavelength not normally absorbed by molecules produced by themicroorganisms. The optical density of the culture samples representsdispersion of the incident beam by the particulate microorganism. Thehigher the optical density value, the greater the amount of microbialgrowth. The optical density results are illustrated in Table 9 below.

TABLE 9 Effect of Nitrogen upon growth of microorganism consortium underaerobic conditions. Optical Density of Culture in Optical DensityOptical Density of Ammonia of Culture with Culture Absent Time MediumTNT Addition of Nitrogen  0 hours 0.006 0.166 0.005  20 hours 0.0080.152 0.018  48 Hours 0.010 0.144 0.023 146 Hours 1.520 0.432 0.073Difference 1.514 0.266 0.068 Over Background NA 3.99 NA

The TNT and No Nitrogen cultures were significantly less productive thanthe ammonia supplemented cultures. Still the TNT supplemented culturevalues were consistently higher than the No Nitrogen values. Thisindicates that the cultures were using TNT as the nitrogen source in theTNT supplemented culture.

EXAMPLE 10

Another study, similar to Example 9 above, was performed under anaerobicconditions. A sample of the 37° C./Log Phase Culture was placed in eachof three fresh media formulations. The first contained mineral saltsdefined medium (MMO) and ammonia as the nitrogen source. The secondcontained MMO and TNT as the nitrogen source. The third contained onlyMMO and no added nitrogen. The cultures were placed in sealed serumbottles and the atmosphere was replaced with pure Nitrogen. Cultureswere incubated without shaking in an incubator at 37° C. The results ofthis study are illustrated in Table 10 below.

TABLE 10 Effect of Nitrogen upon growth of microorganism consortiumunder anaerobic conditions. Optical Density of Culture Optical Densityof in Ammonia Optical Density of Culture Absent Time Medium Culture withTNT Additional Nitrogen  0 hours 0.008 0.204 0.005  20 hours 0.012 0.2180.009  48 Hours 0.017 0.268 0.007 146 Hours 0.482 0.272 0.019 Difference0.474 0.068 0.014 Over NA 4.88  NA background

Once again, the TNT and No Nitrogen cultures were significantly lessproductive than the ammonia supplemented cultures. Still the TNTsupplemented culture values were consistently higher than the NoNitrogen values. This indicates that the cultures were using TNT as thenitrogen source in the TNT supplemented culture. Overall, the anaerobicconditions showed less growth than the aerobic cultures.

EXAMPLE 11

In order to evaluate the thermal resistance of the microorganismconsortium in a system which will adequately mimic those of a pentolitepour, fresh samples of a TNT grown consortium and a control absent TNTwere freeze dried and tested directly for temperature sensitivity. Thefreeze dried samples were placed into aluminum foil packets. Aluminumfoil was used because its heat transference properties ensured that thetemperature experienced by the freeze dried powder approximated thatproduced by the oven. The foil packets were placed in an oven at astarting temperature of either 100° C. or 80° C. The initial 100° C. and80° C. temperatures were maintained for 2 minutes. Each temperature wasthen incrementally decreased at the rate of 1° C. per minute to 35° C.The packets remained at 35° C. for 10 minutes and were then removed fromthe oven. The contents of the packets were placed in MMO with TNT andglycerol, and then placed in a shaking incubator at 37° C.

The negative controls were void of any color which indicates completeabsence of nitrogen degradation. All other samples were in variousstages of TNT degradation as indicated by the color reduction in thesamples from colorless to light orange to deep red or violet. Thesamples that started at 80° C. exhibited more advanced TNT degradationthan those that started at 100° C.

These results were in accordance with the results of Example 8 above. Toreiterate, in that study the samples from the 80° C. water bath had amore optimal survival rate than the samples from the 95° C. water bath.Therefore, although the consortium did respond after experiencingtemperatures as high as 100° C., a maximum of 80° C. represented themore optimal initial temperature.

The lyophilized microorganisms still produced significant bioremediationresults even after being exposed to temperatures corresponding to thatof a hot melt of explosive material. Accordingly, it can be concludedthat lyophilization of microorganisms dramatically improves the thermalresistance of the microorganisms.

EXAMPLE 12

In order to further evaluate the thermal tolerance and protection of themicroorganism consortium, freeze drying was compared withmicroencapsulation. The microencapsulation procedure requiredmaintaining a substantial amount of fresh cell culture. The cells weredivided into 4 samples and resuspended in phosphate buffered saline(PBS), in PBS and 3% dried milk, in PBS and 3% bentonite clay, and inminimal medium with glycerol. Samples of the four suspensions wereprepared by freeze drying 2 ml portions. The remainder of thesuspensions was divided into 2 samples for encapsulation into alginateand polyacrylamide. Encapsulation into alginate was accomplished byadding sodium alginate to the suspension sample and then adding themixture dropwise into a Calcium Chloride solution, with a molarity of0.1. Encapsulation into polyacrylamide was accomplished by combining abiacrylamide mixture with a catalyst, such as a product sold as Temed,and beta-mercaptoethanol. As the mixture polymerized, the microorganismsuspension was trapped in a gel matrix. Half of each sample selected forencapsulation (alginate or polyacrylamide), was freeze dried and theother half was air dried.

All samples (freeze dried, encapsulated and freeze dried, encapsulatedand air dried) were exposed to the temperature curve of Example 8 above.The samples were then added to low temperature agar and overlaid ontotal nutrient agar. Outgrowth and survival of the samples wasevaluated. Additional portions of each sample were then added back tominimal medium with glycerol and TNT to assess the survival of theTNT-critical portions of the consortium.

The encapsulated samples did not result in a significant difference ingrowth as compared with the freeze dried samples. Thus encapsulation didnot offer any distinct advantage over freeze drying with respect totemperature tolerance and subsequent survivability of the microorganismconsortium.

The present invention may be embodied in other specific forms withoutdeparting from its spirit or essential characteristics. The describedembodiments are to be considered in all respects as illustrative onlyand not restrictive. The scope of the invention is, therefore, indicatedby the appended claims rather than by the foregoing description. Allchanges which come within the meaning and range of equivalency of theclaims are to be embraced within their scope.

1. A method for remediating an explosive device, if the explosive deviceonce installed at a predetermined detonation site fails to detonate asintended, said method comprising the steps: (a) forming a quantity of anexplosive material into an explosive device; (b) identifyingmicroorganisms capable of bioremediating said explosive material; (c)shaping a quantity of said microorganisms into an aggregation; (d)positioning said aggregation in such proximity to said quantity of saidexplosive material that, when said quantity of said microorganisms insaid aggregation is mobilized, said microorganisms in said quantitythereof commence bioremediation of said quantity of said explosivematerial; and (e) placing said explosive device at the predetermineddetonation site, whereby if said explosive device fails to detonate asintended, when said quantity of said microorganisms is mobilized, saidmicroorganisms therein deactivate said explosive device bybioremediating said quantity of said explosive material in situ at saiddetonation site.
 2. A method as recited in claim 1, wherein said step offorming occurs before said step of positioning.
 3. A method as recitedin claim 2, wherein said step of positioning comprises the step ofdepositing said aggregation on a surface of said explosive materialexposed in said explosive device.
 4. A method as recited in claim 2,wherein said step of forming comprises the step of disposing saidquantity of said explosive material in a shell, and said shell enableswater from the exterior of said shell to flow into said shell andcontact said quantity of said explosive material disposed therein.
 5. Amethod as recited in claim 4, wherein said step of positioning comprisesthe step of depositing said aggregation on an exposed surface of saidquantity of said explosive material disposed in said shell.
 6. A methodas recited in claim 4, wherein said shell is porous.
 7. A method asrecited in claim 6, wherein said step of positioning comprises the stepof embedding said aggregation in said shell.
 8. A method as recited inclaim 4, wherein said step of positioning comprises the step of locatingsaid aggregation in a recess in an exposed surface of said quantity ofsaid explosive material disposed in said shell.
 9. A method as recitedin claim 8, wherein said step of positioning comprises the step ofembedding said aggregation in said quantity of said explosive materialdeposited in said shell.
 10. A method as recited in claim 4, whereinsaid shell has an open end.
 11. A method as recited in claim 4, whereina hole is formed through said shell.
 12. A method as recited in claim 1,wherein said quantity of said explosive material in said explosivedevice is porous.
 13. A method as recited in claim 1, further comprisingthe step of mixing a surfactant with said quantity of said explosivematerial.
 14. A method as recited in claim 1, wherein said step ofpositioning occurs before said step of forming.
 15. A method as recitedin claim 14, wherein said step of positioning comprises the step ofembedding said aggregation in said quantity of said explosive material.16. A method as recited in claim 14, further comprising the step ofintroducing a thermal protective additive into said aggregation.
 17. Amethod as recited in claim 1, wherein said aggregations have a formselected from the group of forms consisting of a pellet, a capsule, ashard, a flake, a granule, a powder, and a clump.
 18. A method asrecited in claim 1, wherein said microorganisms are among amicroorganism consortion identified at the American Type CultureCollection by ATTCC Designation No.
 55784. 19. A method as recited inclaim 1, further comprising the step of mixing a surfactant with saidquantity of said explosive material.
 20. A method as recited in claim 1,wherein said microorganisms are selected from a group of microorganismsconsisting of Pseudomonas spp., Escherichia Coli, Morganella Morganii,Rhodococcus spp., Comamonas spp., and dentrifying bacteria.
 21. A methodas recited in claim 1, wherein said microorganisms are selected from agroup of microorganisms consisting of microorganisms in Pseudomonas spp.consisting of aeruginosa, fluorescens, acidovorans, mendocina, andcepacia.
 22. A method as recited in claim 1, wherein said explosivematerial is selected from a group of explosive materials consisting oftrinitrotoluene, hexanitrostilbene, hexanitroazobenzene,diaminotrinitrobenzene and triaminotrinitrobenzene, cyclotrimethylenetrinitramine, cyclotetramethylene tetranitramine, nitroguanidine,2,4,6-trinitrophenylmethylnitramine, pentaerythritol tetranitrate,nitroglycerine, and ethylene glycol dinitrate.
 23. A method as recitedin claim 1, further comprising the step of dehydrating said quantity ofsaid microorganisms.
 24. A method as recited in claim 1, furthercomprising the step of freeze drying said quantity of saidmicroorganisms.
 25. A method as recited in claim 1, wherein said step ofpositioning comprises the steps: (a) housing said quantity of saidmicroorganisms in a bioremediation apparatus; and (b) coupling saidbioremediation apparatus to said explosive device.
 26. A method asrecited in claim 25, further comprising the step of shaping saidquantity of said microorganisms into an aggregation having the shape ofa doughnut.
 27. A method as recited in claim 25, further comprising thestep of shaping said quantity of said microorganisms into an aggregationhaving the shape of a block.
 28. A method as recited in claim 1, whereinas a result of said step of forming and said step of positioning, saidaggregation comes to be located in proximity to the portion of saidexplosive device configured to receive a detonator for said explosivedevice.
 29. A method as recited in claim 1, further comprising the stepof introducing a thermal protective additive into said aggregation. 30.A method for remediating an explosive device, if the explosive deviceonce installed at a predetermined detonation site fails to detonate asintended, said method comprising the steps: (a) forming a quantity of anexplosive material into an explosive device; (b) identifyingmicroorganisms capable when mobilized of bioremediating said explosivematerial; (c) housing a quantity of said microorganisms in abioremediation apparatus; (d) coupling said bioremediation apparatus tosaid explosive device; and (e) placing the explosive device with saidbioremediation apparatus coupled thereto at the predetermined detonationsite, whereby if said explosive device fails to detonate as intended,when said quantity of said microorganisms is mobilized, saidmicroorganisms therein deactivate said explosive device bybioremediating said quantity of said explosive material in situ at saiddetonation site.
 31. A method as recited in claim 30, wherein said stepof forming comprises the step of disposing said quantity of saidexplosive material in a shell, and wherein said step of coupling affordsaccess by said quantity of said microorganisms in said bioremediationapparatus to the interior of said shell and said quantity of saidexplosive material disposed therein.
 32. A method as recited in claim31, wherein said shell comprises: (a) an open end, said bioremediationapparatus becoming secured in said open end of said shell in said stepof coupling: (b) a capwell at said open, end of said shell; and (c) abioremediation portal formed through said capwell communicating betweensaid explosive material in said shell and said bioremediation apparatus,when said bioremediation apparatus is coupled to said explosive device.33. A method as recited in claim 30, wherein said bioremediationapparatus comprises: (a) storage means for releasably containing saidquantity of said microorganisms; and (b) divider means for selectivelyreleasing said quantity of said microorganisms from said storage meansinto contact with said quantity of said explosive material in saidexplosive device, when said bioremediation apparatus is coupled to saidexplosive device.
 34. A method as recited in claim 33, wherein saiddivider means comprises a removable barrier.
 35. A method as recited inclaim 34, wherein said barrier is removable mechanically.
 36. A methodas recited in claim 34, wherein said barrier is removable electrically.37. A method as recited in claim 34, wherein said barrier is removablechemically.
 38. A method as recited in claim 33, wherein saidbioremediation apparatus further comprises: (a) reservoir means forreleasably containing a quantity of a liquid capable of mobilizing saidquantity of said microorganisms; and (b) separation means forselectively releasing said quantity of said liquid from said reservoirmeans into said storage means.
 39. A method as recited in claim 30,wherein said bioremediation apparatus comprises: (a) reservoir means forreleasably containing a quantity of a liquid capable of mobilizing saidquantity of said microorganisms; (b) storage means for releasablycontaining said quantity of said microorganisms, said storage meansbeing in selective communication with said reservoir means; (c) firstvalve means for releasing said quantity of said liquid from saidreservoir means into said storage means in an open condition of saidfirst valve means; and (d) second valve means for releasing saidquantity of said microorganisms from said storage means with saidquantity of said liquid into contact with said quantity of saidexplosive material in said explosive device when said bioremediationapparatus is coupled to said explosive device.
 40. A method as recitedin claim 39, wherein said first valve means and said second valve meansare operably interconnected.
 41. A method as recited in claim 30,wherein said microorganisms are selected from a group of microorganismsconsisting of microorganisms in Pseudomonas spp. consisting ofaeruginosa, fluorescens, acidovorans, mendocina, and cepacia.
 42. Amethod as recited in claim 30, further comprising the step of shapingsaid quantity of said microorganisms into an aggregation having theshape of a doughnut.
 43. A method as recited in claim 30, furthercomprising the step of shaping said quantity of said microorganisms intoan aggregation having the shape of a block.
 44. A method as recited inclaim 30, wherein said quantity of said microorganisms is suspended in aliquid.
 45. A method for remediating an explosive device, if theexplosive device once installed at a predetermined detonation site failsto detonate as intended, said method comprising the steps: (a) selectingan explosive material from which to form an explosive device; (b)identifying microorganisms capable of bioremediating said explosivematerial; (c) dispersing a quantity of said microorganisms in a quantityof said explosive material, thereby producing a quantity of an explosivemixture with bioremediating capacity; (d) forming said quantity of saidexplosive mixture into an explosive device; and (e) placing saidexplosive device at the predetermined detonation site, whereby if saidexplosive device fails to detonate as intended, when said quantity ofsaid microorganisms is mobilized, said microorganisms therein deactivatesaid explosive device by bioremediating said quantity of said explosivematerial in situ at said detonation site.
 46. A method as recited inclaim 45, further comprising the step of shaping said quantity of saidmicroorganisms into aggregations.
 47. A method as recited in claim 46,further comprising the step of introducing a thermal protection additiveinto said aggregations.
 48. A method as recited in claim 46, whereinsaid aggregations have a form selected from the group of formsconsisting of a pellet, a capsule, a shard, a flake, a granule, apowder, and a clump.
 49. A method as recited in claim 45, furthercomprising the step of dehydrating said quantity of said microorganism.50. A method as recited in claim 45, further comprising the step offreeze drying said quantity of said microorganisms.
 51. A method asrecited in claim 45, wherein said quantity of said explosive mixture insaid explosive device is porous.
 52. A method as recited in claim 45,wherein said step of forming comprises the step of disposing saidquantity of said explosive mixture in a shell, and said shell enableswater from the exterior of said shell to flow into said shell andcontact said quantity of said explosive mixture disposed therein.
 53. Amethod as recited in claim 52, wherein said shell is porous.
 54. Amethod as recited in claim 52, wherein said shell has an open end.
 55. Amethod as recited in claim 52, wherein a hole is formed through saidshell.
 56. A method as recited in claim 45, wherein said microorganismsare among a microorganism consortium identified at the American TypeCulture Collection by ATCC Designation No.
 55894. 57. A method asrecited in claim 45, further comprising the step of mixing a surfactantwith said quantity of said explosive material.
 58. A method as recitedin claim 45, wherein said microorganisms are selected from a group ofmicroorganisms consisting of Pseudomonas spp., Escherichia Coli,Morganella Morganii, Rhodococcus spp., Comamonas spp., and dentrifyingbacteria.
 59. A method as recited in claim 45, wherein saidmicroorganisms are selected from a group of microorganisms consisting ofmicroorganisms in Pseudomonas spp. consisting of aeruginosa,fluorescens, acidovorans, mendocina, and cepacia.
 60. A method asrecited in claim 45, wherein said explosive material is selected from agroup of explosive materials consisting of organic nitroaromaticexplosives, organic nitramine explosives, and organic nitric esterexplosives.
 61. A method as recited in claim 45, wherein said explosivematerial is selected from a group of explosive materials consisting oftrinitrotoluene, hexanitrostilbene, hexanitroazobenzene,diaminotrinitrobenzene and triaminotrinitrobenzene, cyclotrimethylenetrinitramine, cyclotetramethylene tetranitramine, nitroguanidine,2,4,6-trinitrophenylmethylnitramine, pentaerythritol tetranitrate,nitroglycerine, and ethylene glycol dinitrate.
 62. A method forremediating an explosive device, if the explosive device once installedin a predetermined detonation site fails to detonate as intended, saidmethod comprising the steps: (a) forming a quantity of an explosivematerial into an explosive device; (b) identifying microorganismscapable of bioremediating said explosive material; (c) positioning aquantity of said microorganisms in such proximity to said quantity ofsaid explosive material that, when said microorganisms are mobilized,said microorganisms in said quantity thereof commence bioremediation ofsaid quantity of said explosive material; and (d) placing said explosivedevice at the predetermined detonation site, whereby if said explosivedevice fails to detonate as intended, when said quantity of saidmicroorganisms is mobilized, said microorganisms therein deactivate saidexplosive device by bioremediating said quantity of said explosivematerial in situ at said detonation site.
 63. A method as recited inclaim 62, further comprising the step of shaping said quantity ofmicroorganisms into aggregations.
 64. A method as recited in claim 63,wherein said aggregations have a form selected from the group of formsconsisting of a pellet, a capsule, a shard, a flake, a granule, apowder, and a clump.
 65. A method as recited in claim 62, wherein saidstep of positioning comprises the step of depositing said aggregationson a surface of said explosive material exposed in said explosivedevice.
 66. A method as recited in claim 62, wherein said step offorming comprises the step of disposing said quantity of said explosivematerial in a shell, and said shell enables water from the exterior ofsaid shell to flow into said shell and contact said quantity of saidexplosive material disposed therein.
 67. A method as recited in claim66, wherein said shell is porous, and said step of positioning comprisesthe step of embedding said aggregations in said shell.
 68. A method asrecited in claim 66, wherein said shell has an open end.
 69. A method asrecited in claim 66, wherein a hole is formed through said shell.
 70. Amethod as recited in claim 66, wherein said step of positioningcomprises the step of inserting said aggregations into a recess in anexposed surface of said quantity of said explosive material disposed insaid shell.
 71. A method as recited in claim 62, wherein as a result ofsaid step of forming and said step of positioning, said aggregationscome to be located in proximity to the portion of said explosive deviceconfigured to receive a detonator for said explosive device.
 72. Amethod as recited in claim 62, wherein said step of positioningcomprises the step of dispersing said quantity of said microorganisms insaid quantity of said explosive material, thereby producing a quantityof an explosive mixture with bioremediating capacity.
 73. A method asrecited in claim 62, further comprising the step of dehydrating saidquantity of said microorganisms.
 74. A method as recited in claim 62,further comprising the step of freeze drying said quantity of saidmicroorganisms.
 75. A method as recited in claim 62, further comprisingthe step of mixing a surfactant with said quantity of said explosivematerial.
 76. A method as recited in claim 62, wherein saidmicroorganisms are selected from a group of microorganisms consisting ofPseudomonas spp., Escherichia Coli, Morganella Morganii, Rhodococcusspp., Comamonas spp., and dentrifying bacteria.
 77. A method as recitedin claim 62, wherein said microorganisms are selected from a group ofmicroorganisms consisting of microorganisms in Pseudomonas spp.consisting of aeruginosa, fluorescens, acidovorans, mendocina, andcepacia.